First Fossil Frog from Antarctica: Implications for Eocene High

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OPEN First fossil frog from Antarctica: implications for Eocene high latitude climate conditions and Gondwanan cosmopolitanism of Australobatrachia Thomas Mörs1,2*, Marcelo Reguero3 & Davit Vasilyan4,5

Cenozoic ectothermic continental tetrapods (amphibians and reptiles) have not been documented previously from Antarctica, in contrast to all other continents. Here we report a fossil ilium and an ornamented skull bone that can be attributed to the Recent, South American, anuran family Calyptocephalellidae or helmeted frogs, representing the frst modern amphibian found in Antarctica. The two bone fragments were recovered in Eocene, approximately 40 million years old, sediments on Seymour Island, Antarctic Peninsula. The record of hyperossifed calyptocephalellid frogs outside South America supports Gondwanan cosmopolitanism of the anuran clade Australobatrachia. Our results demonstrate that Eocene freshwater ecosystems in Antarctica provided habitats favourable for ectothermic vertebrates (with mean annual precipitation ≥900 mm, coldest month mean temperature ≥3.75 °C, and warmest month mean temperature ≥13.79 °C), at a time when there were at least ephemeral ice sheets existing on the highlands within the interior of the continent.

Consistent with the geological evidence, it has been hypothesized that the formation of Antarctic ice sheets predates the fnal break-up of Gondwana, the opening of the Drake Passage and the thermal isolation of the continent1–3. Tis is refected by a low diversity of terrestrial mammals on the Antarctic Peninsula during the middle to late Eocene with only two species of large mammals and ten species of small mammals4–6 which sharply contrasts with the highly diverse marine fsh fauna indicating temperate conditions in the Weddell Sea7,8. However, no Cenozoic ectothermic continental vertebrates (freshwater fshes, amphibians and reptiles) have been known from Antarctica so far. Here we report the discovery of a fossil ilium from Seymour Island, Antarctic Peninsula (Fig. 1a,b) which can be assigned to the lissamphibian order Anura, and a fragment of a sculptured skull bone that most probably derived from a hyperossified anuran. We assign the specimens to the South American genus Calyptocephalella. Calyptocephalellids, or helmeted frogs, are widely known from Patagonia since the Late Cretaceous9. Tey became extinct in Argentine Patagonia during the Miocene, probably related to a decrease of humidity caused by the rise of the Andes, since the family survived to the present day in a temperate but humid refuge in the central Chilean Andes10. Te material described here derives from estuarine to marginal-marine deposits of the Eocene La Meseta Formation which were deposited in the James Ross Basin, a back-arc basin east of the Antarctic Peninsula, and which are widely exposed on Seymour Island11,12 (Fig. 1b,c). Te fossil locality IAA 2/95, also known as ‘Marsupial Site’ is a few m2 wide lens of poorly consolidated, shelly conglomerate8,13. It is situated in the central portion of the Cucullaea I Allomember, within unit Telm 5 on the northwestern slope of the mesa (Fig. 1c,d), and informally referred to as the ‘Natica horizon’8,13. It has produced shark, ray and skate teeth, remains of marine bony fshes, as well as teeth of terrestrial mammals, worm (clitellate) cocoons, and seeds of water lilies8,12–22. Based on dinocyst occurrences, the age of this deposit is considered to be about 40 Ma (Bartonian, Eocene)23,24.

1Department of Palaeobiology, Swedish Museum of Natural History, P.O. Box 50007, SE-104 05, Stockholm, Sweden. 2Bolin Centre for Climate Research, Stockholm University, Stockholm, Sweden. 3Instituto Antártico Argentino, Campus Miguelete, 25 de Mayo 1151, 3° piso B1650HMK, San Martín, Buenos Aires, Argentina. 4JURASSICA Museum, Route de Fontenais 21, 2900, Porrentruy, Switzerland. 5Department of Geosciences, University of Fribourg, Chemin du musée 6, 1700, Fribourg, Switzerland. *email: [email protected]

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Figure 1. Maps and stratigraphic column of the studied area and succession. (a) Map of Antarctica showing the location of the study area. (b) Map of the northern Antarctic Peninsula showing the location of Seymour Island. (c) Geological sketch map of Seymour Island, showing the position of the fossil locality (asterisk). (d) Stratigraphic column of the La Meseta Formation on Seymour Island (from11) showing the position of the fossil locality (asterisk). Redrawn from13.

Te fossil frog remains were collected during three joint Argentinian-Swedish expeditions to Seymour Island in the austral summers 2011–13. Te bone fragments were concentrated from dry-sieved sediment samples as described by8,12,20 and sorted by using a Leica MZ6 stereomicroscope. Te material is housed in the palaeozoo- logical collections of the Swedish Museum of Natural History, Stockholm, with the inventory numbers NRM-PZ B281 and B282. Results Systematic palaeontology. Anura Fischer von Waldheim, 181325 Neobatrachia Reig, 195826 Australobatrachia Frost et al., 200627 Calyptocephalellidae Reig, 196028 Calyptocephalella Strand, 192829 Calyptocephalella sp. Figures 2 and 3

Referred specimens. Swedish Museum of Natural History NRM-PZ B282, right ilium (Fig. 2), NRM-PZ B281, skull bone (Fig. 3).

Locality, horizon and age. IAA 2/95, Marsupial site, Seymour Island, Antarctic Peninsula (64°13′58″S; 56°39′06″W). ‘Natica horizon’ within the Cucullaea I Allomember (Telm 5) of the La Meseta Formation, Bartonian (40 Ma), Eocene23,24 (Fig. 1).

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Figure 2. Ilium (NRM-PZ B282) of Calyptocephalella sp. from Seymour Island, Antarctica. Ilium in lateral (a), medial (b), ventral (c) and dorsal (d) views. Magnifed region of the dorsal protuberance in lateral (e – dashed rectangle in orange color) and dorsal (f – dashed rectangle in yellow color) view. Te dashed line in black on (a) indicates the probable outline of the posterior extension of the ventral acetabular expansion. Te asterisk (*) on (a,c) indicates the shallow and broad depression of the ventral acetabular expansion. Te double asterisk (**) indicates the notch caudally from the dorsal protuberance. Te dashed red lines on (e,f) outline the intact bone surface. Abbreviations: ac, acetabulum; ar, acetabular rim; dae, dorsal acetabular expansion; is, iliac shaf; paf, preacetabular fossa; spf, supraacetabular fossa; vae, ventral acetabular expansion; vd, ventral depression.

Measurements. Te preserved part of the ilium measures 3.9 mm in length, the distance from the tip of the dorsal acetabular expansion to the (preserved) tip of the ventral acetabular expansion measures 3.3 mm, the highest height of the acetabular fossa equals 2.5 mm. Te skull bone measures 2.7 mm at its both broadest and longest parts.

Ilium. Te fragmentary right ilium (NRM-PZ B282) lacks the caudal portion of the acetabulum and most of the iliac shaf. Te dorsal acetabular expansion has a smooth lateral surface and is higher than the preserved part of the ventral acetabular expansion (Fig. 2a). A large and deep supraacetabular fossa is present at its base (Fig. 2a,d). Te preserved portion of the acetabulum is concave and its shape allows concluding a (semi-)circular outline. Te acetabular rim is most prominent at its anterior part (Fig. 2a,c). Te barely-developed ventral acetabular expansion projects ventrally. Te posterior-most portion of the ventral acetabular expansion is broken of. However, the anterior portion of the ventral acetabular expansion is higher than the preserved posterior portion. In ventral view (Fig. 2a,c), the lateral surface of the ventral acetabular expansion is convex. Te ventral acetabular expansion possesses a shallow and broad depression. In the preacetabular zone, a small and shallow preacetabular fossa is present (Fig. 2c). Te preserved portion of the iliac shaf is damaged and precludes a confdent statement whether the dorsal protuberance is present or absent. A narrow and shallow longitudinal groove is observable in the lateral surface of the iliac shaf, which probably corresponds to the posterior extension of the ventral

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Figure 3. Skull bone fragment (NRM-PZ B281) of Calyptocephalella sp. from Seymour Island, Antarctica in dorsal (a), ventral (b) and lateral (c) views.

species locality coll nr. SVL (in mm) HT (in mm) RHS (in %) Reference C. pichileufensis Río Pichileufú locality BAR 85b 107 6 5.6 48: Figs. 2 and 4 C. gayi unknown M13105/cas:sua:10082* 54.7 2.9 5.3 65, see Fig. S3 range 5.3–5.6 mean ± SD 5.45 ± 0.21 C. sp. IAA 2/95, Marsupial site NRM-PZ B282 38.4 ± 3.8 2.1 Present work

Table 1. Measurements of the snout-vent length (SVL) and height of the transition from the iliac shaf and ilial body (HT) of some Calyptocephalella spp. and Antarctic ilia (Fig. S3), with the value of the ratio between HT and SVL (RHS = HT/SVL*100%) with the value of the standard deviation (SD). Te reconstructed value of the snout-vent length of the studied ilium is highlighted in bold.

depression (sensu10) (Fig. 2a). However, intact parts of bone surface are preserved slightly ventral to the dorsal margin on both lateral and medial surfaces (Fig. 2e,f). Te one on the lateral surface is a curved shallow groove and runs posteroventrally (Fig. 2e). Tis feature anteroventrally demarcates the slightly elevated roughened scar interpreted above as the dorsal protuberance. Te area between the dorsal acetabular expansion and iliac shaf is slightly projected dorsally. Tis area corresponds to the position of the dorsal protuberance. In fact, no clear evidence of a dorsal protuberance can be found on the ilium, only a slightly roughened area with minimal elevation that corresponds to the dorsal protuberance and the scar for the insertion of the musculus gluteus magnus can be observed. At the caudal side of the dorsal protuberance, a distinct notch is visible (Fig. 2a) which we consider as a further evidence of our interpretation. Te area corresponding to the dorsal protuberance is located anteriorly to the anterior margin of the acetabular rim. Medially, the entire surface opposing the acetabulum is lost and the area preserved more anteriorly is slightly convex medially and smooth. Anteriorly and dorsally, just adjacent to the anterior end of the dorsal protuberance a foramen is present (Fig. 2b). Te fragmentary right ilium can be referred to an anuran based on the following characters30 (the numbers before the characters correspond to the feature numbers of Appendix 1 in Gardner et al.30): 7. (semi-) circular acetabulum; 9. acetabulum with distinct margins; 10. acetabular surface concave; 13. at least dorsal acetabular expansion is strongly divergent; 18. the dorsal protuberance present. Tus, the ilium derives from a small-sized frog (3.8 ± 0.4 cm snout-vent length, see methods, Table 1). Te specimen is partly eroded and rather poorly preserved; however, it can be compared with all South American and Australian frog families (Figs. 4, S1 and S2, Table S1). Te families Ranidae, Bufonidae and Hylidae have not been illustrated in the present work, since their morphology is well known31 (Table S1). Te comparison has been done at family level, since the ilia display diagnostic features characteristic for identifcation of the family (dimensions of the dorsal and ventral acetabular expansions; location of the dorsal protuberance relative to the anterior margin of the acetabular rim etc.32,33). Te studied ilium (NRM-PZ B282) difers in: (1) Reduced anterior portion of the dorsal acetabular expansion from nearly all South American and Australian frog families and the genus Telmatobufo, which have moderately or strongly developed anterior portion of the dorsal acetabular expansion. Only the genus Calyptocephalella (Fig. 4c,e), the families Ranidae31, Pipidae (Fig. S1a), Rhinodermatidae (Fig. S1j), and Leptodactylidae (Fig. S2b) have similar state/morphology of this character. (2) Dorsal protuberance located either at the level of or anteriorly

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Figure 4. 3D models of some skeletal elements of Australobatrachia. (a,f) skull and (b–e,g–j) ilia of Calyptocephalella gayi (a–e); Telmatobufo venustus (f–h); Myobatrachus gouldii (i); Limnodynastes convexiusculus (j). Collection numbers of each specimen are listed in Table S1.

from the anterior margin of the acetabular rim from nearly all families, besides Brachycephalidae (Fig. S1a), Rhinodermatidae (Fig. S1j), Telmatobiidae (Fig. S1k), Hyloididae (Fig. S1m), Leptodactylidae (Fig. S2b) and the genera Calyptocephalella (Fig. 4b–e) and Telmatobufo (Fig. 4g,h). (3) Developed dorsal acetabular expansion from the families Ranidae31, Hylidae31, Bufonidae31, Myobatrachidae (Fig. 4i), Pipidae (Fig. S1i), Microhylidae (Fig. S1b), Telmatobiidae (Fig. S1k), Leptodactylidae (Fig. S2b), Allophrynidae (Fig. S2c), Centrolenidae (Fig. S2d) and the genus Telmatobufo (Fig. 4g,h). Other families have moderately or well-developed dorsal acetabular expansion, however, due to incomplete preservation of the Antarctic frog remain any further comparison is impossible. (4) Weakly developed dorsal protuberance and lack of dorsal tubercle from nearly all families (e.g. Limnodynastidae, Fig. 4j), besides Calyptocephalellidae (Fig. 4b–e,g,h), Myobatrachidae (Fig. 4i), Craugastoridae (Fig. S1e), and Dendrabatidae (Fig. S2e). Among the compared forms, only the South American endemic genus Calyptocephalella resembles all mentioned four characters. In addition to this, a shallow and broad depression on the anterior portion of the ventral acetabular expansion is a unique character observable on our ilium (NRM-PZ B282) and Recent Calyptocephalella gayi (Fig. 4). Further, the fossil ilium displays a ventral depression on its lateral surface anteriorly to the acetabulum (Fig. 2c), a comparable structure can be observed also in the fossil species Calyptocephalella canqueli9 but not in the Recent species C. gayi (Fig. 4).

Skull element. Te second bone fragment (NRM-PZ B281) is fat and slightly curved. Both sides of the bone have diferent structures. One surface is covered by small to large circular or reniform in outline, rather deep pits, which sink in the planar surface of the bone (Fig. 3a). Te diameters of pits vary from 0.1–0.7 mm and some of them are punctured by foramina. Te opposite surface of the bone is in general smooth, slightly deepened and is pierced with some foramina, some of which are preceded by a groove (Fig. 3b). One side of the fragment preserves an unbroken margin of the original bone with a distinct process that is bent and that gives the bone a curved shape (Fig. 3c). Te ornamented surface of the bone projects slightly over this process. Comparable ornamentation, build of pits of diferent size, is found on the dorsal surfaces of diferent cranial and postcranial bones of amphibians and reptiles34,35. Among them, the following groups can be excluded from consideration: (1) Albanerpetontidae (Allocaudata); albanerpetontids are a primary Laurasian lissamphibian group with a sin- gle occurrence in Northern Africa. So far no evidence of a Gondwanan radiation of albanerpetontids exists36. In addition to this, all their ornamented bones (e.g. frontal, premaxillae)37 do not resemble the bone described

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here. (2) Caudata; salamanders are also considered as a Laurasian group, with a number of occurrences in Africa which need critical revision38. In salamanders, ornamented bones are found both among skull bones and on vertebrae (on plates located on the tip of the neural arch)39. Bone ornamentation here (e.g. Tylototriton, Chelotriton, Echinotriton39,40) is represented by a network of pits, ridges and pointy spines that do not resemble the bone described here. (3) Crocodylia; in crocodyliforms, comparable patterns of ornamentation with well-developed pits appear only with growth during later ontogenetic stages41–43. On one hand, the bone dimensions indicate a small-sized animal (corresponding to a juvenile crocodilian without such developed ornamentation). On the other hand, crocodylian osteoderms are fat without any processes, unlike the studied bone. (4) Testudines; shell plates of several turtles, such as Trionyx, Allaeochelys etc.35, are also covered by ornamentation. Te ornamentation is characterized by larger and closely arranged pits, which are not always clearly delimited from each other (see Scheyer35: Fig. 1a). (5) Lacertilia; lizards also have skull bones and osteoderms covered with ornamentation patterns44. Tey all are characterized by a network of spines, grooves, ridges45 and protuberances46, which difers from the morphology on NRM-PZ B281. The ornamentation pattern found in NRM-PZ B281 is comparable to that of some frog genera, i.e. Taumastosaurus47, Beelzebufo48, Calyptocephalella9 and Baurubatrachus49, but only the last three genera are Gondwanan forms and, thus, considered for comparison herein. Beelzebufo is a very large form and the ornamentation pattern is present both on skull bones and on vertebrae50. Calyptocephalella9 and Baurubatrachus49 have very similar ornamentation patterns on the surfaces of hyperossifed skull bones, comparable to our specimen. A recent phylogenetic analysis49 placed the Late Cretaceous Baurubatrachus within both Recent calyptocephalellid genera Calyptocephalella and Telmatobufo. Tough Muzzopappa and Báez10 mention that both Calyptocephalella and Telmatobufo are characterized by a heavily ossifed neurocranium, we can confrm this only for the former genus (Fig. 4a,f). Within Calyptocephalella the ornamentation pattern on skull bones is variable. In C. conquella10, it is built either by network of pits in small individuals, or tuberculated ornamentation in adults. In C. satan9 and C. casamayorensis51, ornamented skull bones are slightly larger than NRM-PZ B281 but they have a similar pattern built of pits. C. pichifeufensis48 is known by larger individuals which show similar ornamentation patterns but with larger pits. In comparison to these species, the Antarctic frog displays an ornamentation most similar to that of C. satan9 and C. casamayorensis48. Taking into account our comparison, we conclude that the ornamented bone fragment NRM-PZ B281 represents a skull bone (most probably a nasal) of a small-sized Calyptocephalella or Baurubatrachus. Given the presence of a small Calyptocephalella as indicated by the ilium in the same, only few m2 measuring outcrop, it is most likely that specimen NRM-PZ B281 belongs to the same genus. A comparable record of an ilium and ornamented bones referable to the genus Calyptocephalella has been mentioned in Báez52. Discussion Among Recent amphibians, the frogs (Anura) have the widest distribution, covering all continents except Antarctica, where the conditions have been uninhabitable for over tens of millions of years. Contrary to all other continents, no traces of any extant amphibian group, all of which belong to the lissamphibian clade, have been documented from Antarctica. Tis paper presents the frst record of a lissamphibian in Antarctica, with Eocene fossils referable to the order Anura, and most likely to the australobatrachian genus Calyptocephalella. Te family Calyptocephalellidae belongs to neobatrachian frogs and is exclusively known from South America53,54. Te fve extant species, including the monospecifc genus Calyptocephalella with hyperossifed skull bones, are restricted to the Chilenean Andes54 while most fossil representatives are known from Argentine Patagonia9,48,53. Today, Calyptocephalella inhabits lowland areas of central Chile (upper elevation limit 500 m) east of the Andes within temperate and humid climates, between latitudes 30–43°S. It has an aquatic or semiaquatic lifestyle and populates standing or slow fowing water bodies (lakes, ponds, streams) in the Valdivian temperate Nothofagus forests54–56. The oldest fossils referable to Calyptocephalella are known from the Upper Cretaceous of Argentina9,52. During the Paleocene–terminal early Miocene, their geographic range was restricted to Patagonia east of the Andes48,51,53,57. Not until the late Pleistocene did they appear west of the Andes, where they have their endemic present-day distribution54–57. Te clade Australobatrachia comprises Myobatrachoidea (families Myobatrachidae + Limnodynastidae sensu27), nowadays distributed in Australia and south of New Guinea, and the family Calyptocephalellidae (Batrachophrynidae sensu27). Australobatrachia are considered as a stem group of the Hyloidea. Te earliest myobatrachoid from Australia is at least as old as early Eocene, based on fragmentary ilia that were referred to the basal extant Lechriodus58. Te split between Calyptocephalellidae and Myobatrachidae (Calyptocephalellidae + Myobatrachoidea sensu27) occurred ~100 Ma (~Early-Late Cretaceous boundary)59. Considering the distributions of extant Australobatrachia (Fig. 5), the earliest fossil records10 and the divergence age (from genetic data)59 of both Calyptocephalellidae and Myobatrachoidea lineages, it is clear that Antarctica had played an important palaeobiogeographic role for Australobatrachia and their consequent dispersal. Because the most recent com- mon ancestor of the clade, including Hyloidae and Myobatrachidae + Calyptocephalellidae, occurred in South America, their origin in South America and consequent dispersal from South America to Australia via Antarctica has been suggested59. Additionally, this suggests one more case of strong faunistic afnities of the continent with South America and Australia4,6,16,60. So far, Antarctica has been considered as a dispersal route, but not as a probable place of origin. Te new fossil fnds support the hypothesis10 that Antarctica may have acted as a center of diversifcation for australobatrachians. Te Seymour Island frog reported herein is the frst vertebrate indicative of freshwater habitats on the Eocene Antarctic Peninsula, following invertebrate and plant evidence12,17 (Fig. 6). It is interesting to note that nearly all fossil localities where Calyptocephalella occurs (excepting those, for which fossil plant data are not available) contain evidence of the presence of Nothofagus, including Seymour Island16,60,61. Te southern extant range of Calyptocephalella occurs sympatrically with the microbiotherian marsupial Dromiciops gliroides (Fig. S4), also known as “Monito del Monte” or “Colocolo Opossum”, a small mammal with an arboreal lifestyle and an endemic

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Figure 5. Eocene palaeogeography of the south polar region with a cladogram of australobatrachid frogs showing their occurrences on the southern continents. Te grey color indicates the outlines of the continents during the Eocene, the black colored outline the present-day outlines of the continents. Map redrawn from an original generated using ArcGIS 10.17.1 (www.esri.com) sofware, based on the Satellite base map layer in google Maps (Map data ©2019 Google). Abbreviations: ANT, Antarctica; AUS, Australia and Tasmania; NG, New Guinea; NZ, New Zealand; SA, South America. Te red star indicates the fossil locality on Seymour Island.

distribution in the dense Valdivian Nothofagus forests of highland Argentina and Chile62. Te climate of this Nothofagus forest area with the sympatric occurrences of these two endemic animals shows humid and temperate conditions (for the numerical values, see Methods and Table 2, Fig. S4). Dromiciops gliroides is the only extant species of the order Microbiotheria and is considered as the only South American representative of the superor- der Australidelphia which otherwise comprises Australian marsupials63. From the same small shell-rich lens that produced the frog remains reported herein, the fossil microbiotherian Woodburnodon casei has been described64. Hence, we hypothesize that the climatic conditions for the Antarctic Peninsula during the Bartonian (late middle Eocene) should be comparable with the climate found today in the concurrent range of the Calyptocephalella- and Dromiciops-inhabited Nothofagus forests of South America. Te fossil fnds of a frog and marsupial from Seymour Island, and their fossil and Recent distributions, repre- sent outstanding examples of the role of global climate change on shifing biogeographic ranges. Despite global cooling and the disappearance of the habitats of these groups over large areas from Antarctica to Patagonia, they maintained their relictual occurrence in the Nothofagus forests of the central Chilean Andes. Tus, the Valdivian Nothofagus forest is a unique environment ofering habitats not only for Eocene Antarctic refugees but also provides a modern analogue of the Antarctic climate just prior to the glaciation of the southern continent. Methods Documentation. Te studied fossil material has been mounted on aluminum stubs, coated with gold and imaged using a Hitachi S-4300 feld emission scanning-electron microscope at the Swedish Museum of Natural History (Stockholm). For the comparison with relevant families (Table S1), the CT data from the website Morphosource65 have been used. Te visualization and segmentation of the bone material have been performed using the Amira 9.0 sofware in Porrentruy, Switzerland.

Nomenclature. If not otherwise indicated, the osteological nomenclature of this study follows that of Gómez and Turazzini66 for the description of the fossil remains. Body size estimation. Te values of the snout-vent length (SVL) of Calyptocephalella sp. from Antarctica have been reconstructed using photographs of the skeleton of C. pichileufensis48 and the 3D model of C. gayi

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Figure 6. Reconstruction of an Eocene pond in the Nothofagus forest of the Antarctic Peninsula with Calyptocephalella, sitting on a leaf of Notonuphar antarctica which was described from the same locality12. Artwork by Pollyanna von Knorring, Swedish Museum of Natural History. Photo credits: Simon Pierre Barrette and José Grau de Puerto Montt, Wikimedia Commons (CC BY-SA 3.0), and Mats Wedin, Swedish Museum of Natural History.

Station number 1 2 3 4 5 6 7 8 9 City Loanco Nirivilo Chillan Antuco Curacautin Choshuenco Huequi Puerto Montt Valdivia Elevation (m) 70 220 127 550 541 200 30 100 100 MAP (mm) 877.82 919.5 998 1024.5 988.7 1903.53 1830 1610.8 1870 MCMT (°C) 8.74 7.9 7.69 5.27 3.75 4.51 5.04 5.57 6.9 MWMT (°C) 18.05 18.04 18.45 17.28 15.5 15 13.79 14.7 15.5

Table 2. Climatic parameters (MAP, MCMT, MWMT) and the elevation at the climatic stations according to the database of the word bank groups67. Abbreviations: MAP = mean annual precipitation, MCMT = mean coldest month temperature, MWMT = mean warmest month temperature. Te location of the climatic stations can be found on Fig. S4.

(Table 1, Fig. S3). Te height of the transition (HT) from the iliac shaf and ilial body (Fig. S3) have been used as reference for comparison to reconstruct the body size of NRM-PZ B282. Te ratio of the HT to SVL has been used as a reference to calculate the value of the snout-vent lenth of the individual NRM-PZ B282.

Climatic analysis. We analyzed the climatic parameters of selected stations from the area with sympatric occurrence of Dromiciops gliroides and Calyptocephalella gayi (Table 2 and Fig. S4). Since the upper elevation limit for extant Calyptocephalella distribution is 500 m above sea level, only stations up to this elevation have been considered for climate analysis. Tis analysis shows a remarkable climatic space with mean annual precipitation ≥ 900 mm, coldest month mean temperature ≥ 3.75 °C, and warmest month mean temperature ≥ 13.79 °C (Table 2). Te climatic parameter variations result from both elevation and latitudinal diferences, so the temperature increases and the precipitation decreases northwards, whereas an opposite trend is observable at higher altitudes.

Received: 8 November 2019; Accepted: 4 March 2020; Published: xx xx xxxx

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Worthy and an anonymous reviewer for their careful work which improved the manuscript signifcantly. Tis work was funded by the Swedish Research Council (VR grant number 2009-4447) to T.M., the Bolin Center for Climate Research, Stockholm University (RA6 grant) to T.M., the Consejo Nacional de lnvestigaciones Cientifcas y Tecnicas (CONICET grant number PIP 0462) to M.R., and the Argentinian National Agency for Promotion of Science and Technology (ANPCyT grant number PICTO 0093/2010) to M.R. Author contributions T.M. and M.R. collected and processed the material. D.V. and T.M. analyzed the material. D.V. photographed the material, performed 3D reconstructions and provided the climate data. All authors discussed the results, wrote the manuscript and provided illustrations. Competing interests Te authors declare no competing interests.

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